The effect of titanium carbide used in implants on stemness and senescence of mouse bone marrow derived mesenchymal stem cells

Abstract

Background: The aim of the present study was to determine the effect of Titanium carbide nanoparticles on stemness and senescence markers of mouse bone marrow derived mesenchymal stem cells. Methods: After isolation of mesenchymal stem cells from mouse’s bone marrow, their surface markers were studied using flowcytometry. MTT assay, cell cycle analysis; colony forming assay, and senescent beta galactosidase staining were performed for cells treated with titanium carbide nanoparticles. Then, the expression of OCT-4 and Nanog genes were studied by qRT-PCR in titanium carbide treated mouse’s bone marrow mesenchymal stem cells (mBMSCs). Results: All mBMSCs showed spindle shaped morphology. Their identity was confirmed by flow cytometry of stained cells for CD11b, CD90, CD45, and CD44 markers. The MTT assay results showed that titanium carbide effect is time-dependent and no cytotoxic effects were observed in short treatment times. But toxic effects were observed after 72 or 144 hours of post treatment with doses range from 0.1 to 1 mM (P<0.05). Besides, cell cycle study detected more cell populations in G0/G1 and less percentage in S phase. The colony forming assay in treated cells exhibited smaller size of colonies. The beta galactosidase staining of treated cells demonstrated more positive cells (more aged cells). Finally, qRT-PCR showed significant down regulation of OCT-4 and Nanog (p<0.05). Conclusion: Titanium carbide as an implant material could affect the viability, stemness and senescence of mBMSCs in negative manner.